US4108794A - Oil immersion liquid for fluorescence microscopes - Google Patents

Oil immersion liquid for fluorescence microscopes Download PDF

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Publication number
US4108794A
US4108794A US05/655,949 US65594976A US4108794A US 4108794 A US4108794 A US 4108794A US 65594976 A US65594976 A US 65594976A US 4108794 A US4108794 A US 4108794A
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United States
Prior art keywords
oil
oil immersion
fluorescence
immersion liquid
cargile
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Expired - Lifetime
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US05/655,949
Inventor
Ken Yonekubo
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Olympus Corp
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Olympus Optical Co Ltd
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    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/33Immersion oils, or microscope systems or objectives for use with immersion fluids

Definitions

  • the present invention relates to an oil immersion liquid for use with fluorescence microscopes.
  • cargile oil and cedar oil are used as oil immersion liquids for ordinary microscopes but these oils emit auto fluorescence.
  • distilled water which does not emit auto fluorescence glycerin which emits weak auto fluorescence but scarcely affects fluorescence microscopy and cargile oil which emits relatively weak auto fluorescence are also used as oil immersion liquids for fluorescence microscopy.
  • cargile oil emits certain degrees of auto fluorescence which unavoidably produces flare and affects fluorescence microscopic observations. Distilled water does not emit auto fluorescence and is deal for fluorescence microscopy in this respect.
  • Glycerin scarcely affects fluorescence microscopy with its auto fluorescence as described above and has a refractive index of 1.458 which is close to that of cargile oil, therefore being usable with objectives designed for use with cargile oil simply by slightly modifying the objectives.
  • glycerin is moisture-absorptive and its refractive index decreases close down to that of distilled water as time elapses. This decrease in refractive index of glycerin degrades both resolution and image contrast similarly to the case where distilled water is used as an oil immersion liquid.
  • a general object of the present invention is to provide an oil immersion liquid for fluorescence microscopes which consists of silicone oil and permits favorably observing images of fluorescent specimens.
  • silicone oil When silicone oil is adopted as an oil immersion liquid for fluorescence microscopes, it emits no auto fluorescence like distilled water and, therefore, brings about no degradation in resolution or image contrast due to its auto fluorescence. Further, it has a refractive index close to that of cargile oil and therefore permits favorably observing images of fluorescent specimens without designing special objectives for fluorescence microscopes. For such fluorescence microscopy by using silicone oil as an oil immersion liquid, it will be sufficient to slightly adjust the conventionally designed objectives, for example, by slightly displacing some lens components of the objectives along their optical axes. Furthermore, silicone oil used as an oil immersion liquid is free from refractive index variation which degrade resolution and image contrast since silicone oil is not moisture-absorptive or volatile.
  • the present invention provides an excellent oil immersion liquid which is free from auto fluorescence and variation in refractive index by adopting silicone oil as an oil immersion liquid for fluorescence microscopes.

Abstract

An oil immersion liquid consisting of silicone oil for fluorescence microscopes which does not emit any auto fluorescence and therefore permits observing microscopic images of fluorescent substances with high contrast when such specimens are observed through oil immersion type objectives using this oil immersion liquid.

Description

BACKGROUND OF THE INVENTION
a. Field of the invention
The present invention relates to an oil immersion liquid for use with fluorescence microscopes.
B. Description of the prior art
Objectives designed for a magnification of x100 and a numerical aperture N.A. of 1.25 or higher are required for studying chromosomes, etc. by using fluorescence microscopes. For this purpose, oil immersion type objectives are adopted for this type of fluorescence microscopes. However, fluorescence emitted from specimens to be observed is generally at a low intensity and satisfactory observation is impossible when images of specimens to be observed are not formed with high contrast. It is, therefore, impossible to satisfactorily observe images of fluorescent specimens when image contrast is degraded due to flare produced by the lens system or auto fluorescence emitted from a substance existing in the lens system or related components.
Conventionally, cargile oil and cedar oil are used as oil immersion liquids for ordinary microscopes but these oils emit auto fluorescence.
Further, distilled water which does not emit auto fluorescence, glycerin which emits weak auto fluorescence but scarcely affects fluorescence microscopy and cargile oil which emits relatively weak auto fluorescence are also used as oil immersion liquids for fluorescence microscopy. Of the above-mentioned oil immersion liquids, cargile oil emits certain degrees of auto fluorescence which unavoidably produces flare and affects fluorescence microscopic observations. Distilled water does not emit auto fluorescence and is deal for fluorescence microscopy in this respect. However, aberrations are corrected for cargile oil in general oil immersion type objectives, while distilled water has a refractive index (Nd = 1.333) which is largely different from that of cargile oil (Nd = 1.514). Therefore, distilled water will generally aggravates aberrations and, in addition, degrades resolution and image contrast, making it impossible to satisfactorily observe images of fluorescent specimens. As a result, it is required to use an objective specially designed for use with distilled water in order to use it as an oil immersion liquid for fluorescence microscopy. In addition, distilled water has another drawback that it has a surface tension which is too low to select a long working distance when it is used as an oil immersion liquid. Glycerin scarcely affects fluorescence microscopy with its auto fluorescence as described above and has a refractive index of 1.458 which is close to that of cargile oil, therefore being usable with objectives designed for use with cargile oil simply by slightly modifying the objectives. However, glycerin is moisture-absorptive and its refractive index decreases close down to that of distilled water as time elapses. This decrease in refractive index of glycerin degrades both resolution and image contrast similarly to the case where distilled water is used as an oil immersion liquid.
SUMMARY OF THE INVENTION
A general object of the present invention is to provide an oil immersion liquid for fluorescence microscopes which consists of silicone oil and permits favorably observing images of fluorescent specimens.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
The present invention has been achieved by paying attention to the fact that silicone oil does not emit auto fluorescence at all and has a refractive index Nd = 1.40 which is relatively close to that of cargile oil. That is to say, the oil immersion liquid according to the present invention is available by using silicone oil as an oil immersion liquid for fluorescence microscopes.
When silicone oil is adopted as an oil immersion liquid for fluorescence microscopes, it emits no auto fluorescence like distilled water and, therefore, brings about no degradation in resolution or image contrast due to its auto fluorescence. Further, it has a refractive index close to that of cargile oil and therefore permits favorably observing images of fluorescent specimens without designing special objectives for fluorescence microscopes. For such fluorescence microscopy by using silicone oil as an oil immersion liquid, it will be sufficient to slightly adjust the conventionally designed objectives, for example, by slightly displacing some lens components of the objectives along their optical axes. Furthermore, silicone oil used as an oil immersion liquid is free from refractive index variation which degrade resolution and image contrast since silicone oil is not moisture-absorptive or volatile. Silicone oil has viscosity Cs = 1.000 to 3,000 which is on the order similar to that of cargile oil and refractive index Nd = 1.40 which is relatively close to that of cargile oil. Therefore, silicone oil allows to select long working distances for objectives of fluorescence microscopes and can be used with general oil immersion type objectives designed for use with cargile oil through simple modification, for example, by slightly displacing the second lens component of the object system along its optical axis.
As is easily understood from the foregoing descriptions, the present invention provides an excellent oil immersion liquid which is free from auto fluorescence and variation in refractive index by adopting silicone oil as an oil immersion liquid for fluorescence microscopes.

Claims (2)

I claim:
1. In a process for using a fluorescent microscope for studying materials which fluoresce in which an oil immersion objective is employed, the improvement consisting of using silicone oil having a refractive index Nd = 1.40 as the oil immersion liquid.
2. The process of claim 1, wherein said silicone oil has a viscosity Cs = 1,000 to 3,000.
US05/655,949 1975-02-14 1976-02-06 Oil immersion liquid for fluorescence microscopes Expired - Lifetime US4108794A (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP50-18025 1975-02-14
JP1802575A JPS5510172B2 (en) 1975-02-14 1975-02-14

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JP (1) JPS5510172B2 (en)
DE (1) DE2606064C3 (en)

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4465621A (en) * 1983-07-01 1984-08-14 R. P. Cargille Laboratories, Inc. Immersion oil for microscopy and related applications
US4491533A (en) * 1983-09-08 1985-01-01 R. P. Cargille Laboratories, Inc. Immersion oil for fluorescence microscopy
US4493533A (en) * 1981-04-27 1985-01-15 Hoffmann-La Roche Inc. Immersion oil containing aliphatic thio compounds
US4559147A (en) * 1982-12-14 1985-12-17 Hoffmann-La Roche Inc. Optical immersion oil
US4587042A (en) * 1984-02-27 1986-05-06 Michael Liva Immersion oil system
US6191852B1 (en) 1997-10-14 2001-02-20 Bayer Aktiengesellschaft Optical measurement system for detecting luminescence or fluorescence signals
US6221281B1 (en) 1998-11-05 2001-04-24 Nikon Corporation Liquid immersion oil
US20040202757A1 (en) * 1999-08-14 2004-10-14 The Procter & Gamble Company Methods and systems utilizing delayed dilution, mixing, and filtration for providing customized beverages on demand
US20040220070A1 (en) * 2003-02-27 2004-11-04 Hans-Joachim Weippert Immersion liquid for water-immersion microscopy
US20040240047A1 (en) * 2003-02-21 2004-12-02 Shafer David R. Catadioptric imaging system for broad band microscopy
US20050152027A1 (en) * 2003-02-21 2005-07-14 Armstrong J. J. Catadioptric imaging system employing immersion liquid for use in broad band microscopy
WO2007140012A2 (en) * 2006-05-26 2007-12-06 Massachusetts Institute Of Technology Immersion fluids for lithography
US20080186471A1 (en) * 2007-02-06 2008-08-07 C8 Medisensors Inc. Apparatus for Stabilizing Mechanical, Thermal, and Optical Properties and for Reducing the Fluorescence of Biological Samples for Optical Evaluation
US20080247036A1 (en) * 2003-02-21 2008-10-09 Armstrong J Joseph Catadioptric microscope objective employing immersion liquid for use in broad band microscopy
US20130057953A1 (en) * 2011-09-02 2013-03-07 Olympus Corporation Nonlinear optical microscope
US8411265B2 (en) 2011-06-14 2013-04-02 C8 Medisensors Inc. Apparatus for stabilizing mechanical, thermal, and optical properties and for reducing the fluorescence of biological samples for optical evaluation
CN106772973A (en) * 2016-12-19 2017-05-31 中国石油天然气股份有限公司 A kind of optical microscope method and device

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5510172B2 (en) * 1975-02-14 1980-03-14
JPH036716U (en) * 1988-11-02 1991-01-23
US5436766A (en) * 1992-09-04 1995-07-25 Lockheed Missiles & Space Company, Inc. Bond between a rigid refractive element and a surrounding housing structure in an optical system containing a liquid refractive element
JP4709505B2 (en) * 2004-06-23 2011-06-22 オリンパス株式会社 Optical device for fluorescence observation
JP2007034020A (en) * 2005-07-28 2007-02-08 Olympus Corp Microscope objective lens

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US2978298A (en) * 1956-09-04 1961-04-04 Gen Electric Process for producing hydrophobic aerogels
US3297393A (en) * 1963-09-30 1967-01-10 Bausch & Lomb Oil immersion type microscope objective
DE1472294A1 (en) * 1966-09-07 1969-02-13 Leitz Ernst Gmbh Immersion body
US3437402A (en) * 1966-02-23 1969-04-08 Bausch & Lomb Aryl thioether immersion liquids
US3497562A (en) * 1966-02-23 1970-02-24 Bausch & Lomb 2,3'-bis(phenylthio)diphenyl sulfide
US3639260A (en) * 1969-04-16 1972-02-01 Nalco Chemical Co Method of preparing antifoaming agent
US3876288A (en) * 1972-09-08 1975-04-08 West Electric Co Light controlling device
US3929667A (en) * 1974-06-19 1975-12-30 R P Cargille Lab Inc Immersion oil formulations for use in microscopy and similar fields
DE2606064A1 (en) * 1975-02-14 1976-09-02 Olympus Optical Co IMMERSION LIQUID
US3979301A (en) * 1975-06-17 1976-09-07 Idemitsu Petrochemical Co., Ltd. Immersion oil for microscopy

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DE677870C (en) * 1937-10-01 1939-07-04 Zeiss Carl Fa Process for the production of immersion oil for microscopic purposes
DE809719C (en) * 1948-12-01 1951-08-02 Bayer Ag Immersion liquid
DE904353C (en) * 1952-03-09 1954-02-18 Bosch Gmbh Robert Method and device for examining the surface quality of bodies
US3434776A (en) * 1965-07-29 1969-03-25 Dow Corp Laser communication system using flexible silicone transmission line
GB1259383A (en) * 1969-03-13 1972-01-05
FR2077731A1 (en) * 1970-02-10 1971-11-05 Auzel Francois
US3655274A (en) * 1970-05-07 1972-04-11 Ingenuics Inc Gravity operated liquid prism

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2978298A (en) * 1956-09-04 1961-04-04 Gen Electric Process for producing hydrophobic aerogels
US3297393A (en) * 1963-09-30 1967-01-10 Bausch & Lomb Oil immersion type microscope objective
US3437402A (en) * 1966-02-23 1969-04-08 Bausch & Lomb Aryl thioether immersion liquids
US3497562A (en) * 1966-02-23 1970-02-24 Bausch & Lomb 2,3'-bis(phenylthio)diphenyl sulfide
DE1472294A1 (en) * 1966-09-07 1969-02-13 Leitz Ernst Gmbh Immersion body
US3639260A (en) * 1969-04-16 1972-02-01 Nalco Chemical Co Method of preparing antifoaming agent
US3876288A (en) * 1972-09-08 1975-04-08 West Electric Co Light controlling device
US3929667A (en) * 1974-06-19 1975-12-30 R P Cargille Lab Inc Immersion oil formulations for use in microscopy and similar fields
DE2606064A1 (en) * 1975-02-14 1976-09-02 Olympus Optical Co IMMERSION LIQUID
US3979301A (en) * 1975-06-17 1976-09-07 Idemitsu Petrochemical Co., Ltd. Immersion oil for microscopy

Cited By (30)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4493533A (en) * 1981-04-27 1985-01-15 Hoffmann-La Roche Inc. Immersion oil containing aliphatic thio compounds
US4559147A (en) * 1982-12-14 1985-12-17 Hoffmann-La Roche Inc. Optical immersion oil
US4465621A (en) * 1983-07-01 1984-08-14 R. P. Cargille Laboratories, Inc. Immersion oil for microscopy and related applications
US4491533A (en) * 1983-09-08 1985-01-01 R. P. Cargille Laboratories, Inc. Immersion oil for fluorescence microscopy
US4587042A (en) * 1984-02-27 1986-05-06 Michael Liva Immersion oil system
US6191852B1 (en) 1997-10-14 2001-02-20 Bayer Aktiengesellschaft Optical measurement system for detecting luminescence or fluorescence signals
US6221281B1 (en) 1998-11-05 2001-04-24 Nikon Corporation Liquid immersion oil
US20040202757A1 (en) * 1999-08-14 2004-10-14 The Procter & Gamble Company Methods and systems utilizing delayed dilution, mixing, and filtration for providing customized beverages on demand
US7307783B2 (en) * 2003-02-21 2007-12-11 Kla-Tencor Technologies Corporation Catadioptric imaging system employing immersion liquid for use in broad band microscopy
US20080247035A1 (en) * 2003-02-21 2008-10-09 Kla-Tencor Technologies Corporation Catadioptric imaging system employing immersion liquid for use in broad band microscopy
US20050152027A1 (en) * 2003-02-21 2005-07-14 Armstrong J. J. Catadioptric imaging system employing immersion liquid for use in broad band microscopy
US8675276B2 (en) * 2003-02-21 2014-03-18 Kla-Tencor Corporation Catadioptric imaging system for broad band microscopy
US7884998B2 (en) * 2003-02-21 2011-02-08 Kla - Tencor Corporation Catadioptric microscope objective employing immersion liquid for use in broad band microscopy
US7633675B2 (en) 2003-02-21 2009-12-15 Kla-Tencor Technologies Corporation Catadioptric imaging system employing immersion liquid for use in broad band microscopy
US20040240047A1 (en) * 2003-02-21 2004-12-02 Shafer David R. Catadioptric imaging system for broad band microscopy
US20080247036A1 (en) * 2003-02-21 2008-10-09 Armstrong J Joseph Catadioptric microscope objective employing immersion liquid for use in broad band microscopy
US20040220070A1 (en) * 2003-02-27 2004-11-04 Hans-Joachim Weippert Immersion liquid for water-immersion microscopy
US7285231B2 (en) * 2003-02-27 2007-10-23 Carl-Zeiss-Stiftung Trading As Carl Zeiss Immersion liquid for water-immersion microscopy
WO2005094304A3 (en) * 2004-03-29 2006-12-21 Kla Tencor Tech Corp Catadioptric imaging system employing immersion liquid for use in broad band microscopy
US7745102B2 (en) 2006-05-26 2010-06-29 Massachusetts Institute Of Technology Immersion fluids for lithography
WO2007140012A3 (en) * 2006-05-26 2008-09-18 Massachusetts Inst Technology Immersion fluids for lithography
US20080063989A1 (en) * 2006-05-26 2008-03-13 Mass Institute Of Technology Immersion fluids for lithography
WO2007140012A2 (en) * 2006-05-26 2007-12-06 Massachusetts Institute Of Technology Immersion fluids for lithography
US20080186471A1 (en) * 2007-02-06 2008-08-07 C8 Medisensors Inc. Apparatus for Stabilizing Mechanical, Thermal, and Optical Properties and for Reducing the Fluorescence of Biological Samples for Optical Evaluation
WO2008098049A2 (en) * 2007-02-06 2008-08-14 C8 Medisensors Inc. Apparatus for stabilizing mechanical, thermal, and optical properties and for reducing the fluorescence of biological samples for optical evaluation
US7973925B2 (en) 2007-02-06 2011-07-05 C8 Medisensors Inc. Apparatus for stabilizing mechanical, thermal, and optical properties and for reducing the fluorescence of biological samples for optical evaluation
WO2008098049A3 (en) * 2007-02-06 2008-09-25 C8 Medisensors Inc Apparatus for stabilizing mechanical, thermal, and optical properties and for reducing the fluorescence of biological samples for optical evaluation
US8411265B2 (en) 2011-06-14 2013-04-02 C8 Medisensors Inc. Apparatus for stabilizing mechanical, thermal, and optical properties and for reducing the fluorescence of biological samples for optical evaluation
US20130057953A1 (en) * 2011-09-02 2013-03-07 Olympus Corporation Nonlinear optical microscope
CN106772973A (en) * 2016-12-19 2017-05-31 中国石油天然气股份有限公司 A kind of optical microscope method and device

Also Published As

Publication number Publication date
JPS5193239A (en) 1976-08-16
DE2606064B2 (en) 1977-04-07
JPS5510172B2 (en) 1980-03-14
DE2606064A1 (en) 1976-09-02
DE2606064C3 (en) 1981-08-06

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